Two pHZ1358 Derivative Vectors for Efficient Gene Knockout in Streptomyces
Two pHZ1358 Derivative Vectors for Efficient Gene Knockout in Streptomyces
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两种用于链霉菌中高效基因敲除的 pHZ1358 衍生载体
DOI：
10.4014/jmb.0910.10031
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发表时间：
2010-04-01
影响因子：
2.8
通讯作者：
Deng, Zixin
中科院分区：
文献类型：
--
作者：
He Yunlong;Wang, Zhijun;Deng, Zixin
链霉菌pIJ101中STI基因的缺失使其衍生物PHZ1358成为基因中断和替换的有效载体。在此基础上，通过构建衍生载体pJTU1278进一步优化了pHZ1358，其中引入了携带多个克隆位点的盒和LacZ选择标记，以方便在大肠杆菌中构建载体。此外，pJTU1278的ORIT区也被删除，产生了可用于PCR靶向的载体(PJTU1289)。阿维链霉菌中阿维菌素生物合成相关基因的缺失证明了这些载体的有效利用。
The deletion of sti from the Streptomyces plasmid pIJ101 made its derivative pHZ1358 an efficient vector for gene disruption and replacement. Here, pHZ1358 was further optimized by the construction of a derivative plasmid pJTU1278, in which a cassette carrying multiple cloning sites and a lacZ selection marker were introduced for convenient plasmid construction in E. coli. In addition, the oriT region of pJTU1278 was also deleted, generating a vector (pJTU1289) that can be used specifically for PCR-targeting. The efficient usage of these vectors was demonstrated by the deletion of the gene involved in avermectin biosynthesis in S. avermitilis.